Aryl ureas as kinase inhibitors

ABSTRACT

This invention relates to new aryl ureas and methods for their synthesis. The inventive compounds are useful in the treatment of (i) raf mediated diseases, for example, cancer, (ii) p38 mediated diseases such as inflammation and osteoporosis, and (iii) VEGF mediated diseases such as angiogenesis disorders.

FIELD OF THE INVENTION

[0001] This invention relates to aryl ureas and methods for theirsynthesis. The inventive compounds are useful in the treatment of

[0002] (i) raf mediated diseases, for example, cancer,

[0003] (ii) p38 mediated diseases such as inflammation and osteoporosis,and

[0004] (iii) VEGF mediated diseases such as angiogenesis disorders.

BACKGROUND OF THE INVENTION

[0005] Activation of the Ras signal transduction pathway indicates acascade of events that have a profound impact on cellular proliferation,differentiation, and transformation. Raf kinase, a downstream effectorof Ras, is a key mediator of these signals from cell surface receptorsto the cell nucleus (Lowy, D. R.; Willumsen, B. M. Ann. Rev. Biochem.1993, 62, 851; Bos, J. L. Cancer Res. 1989, 49, 4682). It has been shownthat inhibiting the effect of active ras by inhibiting the raf kinasesignaling pathway by administration of deactivating antibodies to rafkinase or by co-expression of dominant negative raf kinase or dominantnegative MEK, the substrate of raf kinase, leads to the reversion oftransformed cells to the normal growth phenotype (see: Daum et al.Trends Biochem. Sci. 1994, 19, 474-80; Fridman et al. J. Biol. Chem.1994, 269, 30105-8. Kolch et al. (Nature 1991, 349, 426-28) have furtherindicated that inhibition of raf expression by antisense RNA blocks cellproliferation in membrane-associated oncogenes. Similarly, inhibition ofraf kinase (by antisense oligodeoxynucleotides) has been correlated invitro and in vivo with inhibition of the growth of a variety of humantumor types (Monia et al., Nat. Med. 1996, 2, 668-75). Thus, smallmolecule inhibitors of Raf kinase activity are important agents for thetreatment of cancer (Naumann, U.; Eisenmann-Tappe, I.; Rapp, U. R.Recent Results Cancer Res. 1997, 143, 237; Monia, B. P.; Johnston, J.F.; Geiger, T.; Muller, M.; Fabbro, D. Nature Medicine 1996, 2, 668).

[0006] Inhibition of p38 has been shown to inhibit both cytokineproduction (eg., TNFα, IL-1, IL-6, IL-8) and proteolytic enzymeproduction (eg., MMP-1, MMP-3) in vitro and/or in vivo. The mitogenactivated protein (MAP) kinase p38 is involved in IL-1 and TNF signalingpathways (Lee, J. C.; Laydon, J. T.; McDonnell, P. C.; Gallagher, T. F.;Kumar, S.; Green, D.; McNulty, D.; Blumenthal, M. J.; Heys, J. R.;Landvatter, S. W.; Stricker, J. E.; McLaughlin, M. M.; Siemens, I. R.;Fisher, S. M.; Livi, G. P.; White, J. R.; Adams, J. L.; Yound, P. R.Nature 1994, 372, 739).

[0007] Clinical studies have linked TNFα production and/or signaling toa number of diseases including rheumatoid arthritis (Maini. J. RoyalColl. Physicians London 1996, 30, 344). In addition, excessive levels ofTNFα have been implicated in a wide variety of inflammatory and/orimmunomodulatory diseases, including acute rheumatic fever (Yegin et al.Lancet 1997, 349, 170), bone resorption (Pacifici et al. J. Clin.Endocrinol. Metabol. 1997, 82, 29), postmenopausal osteoperosis(Pacifici et al. J. Bone Mineral Res. 1996, 11, 1043), sepsis (Blackwellet al. Br. J. Anaesth. 1996, 77, 110), gram negative sepsis (Debets etal. Prog. Clin. Biol. Res. 1989, 308, 463), septic shock (Tracey et al.Nature 1987, 330, 662; Girardin et al. New England J. Med. 1988, 319,397), endotoxic shock (Beutler et al. Science 1985, 229, 869; Ashkenasiet al. Proc. Nat'l. Acad. Sci. USA 1991, 88, 10535), toxic shocksyndrome, (Saha et al. J. Immunol. 1996, 157, 3869; Lina et al. FEMSImmunol. Med. Microbiol. 1996, 13, 81), systemic inflammatory responsesyndrome (Anon. Crit. Care Med. 1992, 20, 864), inflammatory boweldiseases (Stokkers et al. J. Inflamm. 1995-6, 47, 97) including Crohn'sdisease (van Deventer et al. Aliment. Pharmacol. Therapeu. 1996, 10(Suppl. 2), 107; van Dullemen et al. Gastroenterology 1995, 109, 129)and ulcerative colitis (Masuda et al. J. Clin. Lab. Immunol. 1995, 46,111), Jarisch-Herxheimer reactions (Fekade et al. New England J. Med.1996, 335, 311), asthma (Amrani et al. Rev. Malad. Respir. 1996, 13,539), adult respiratory distress syndrome (Roten et al. Am. Rev. Respir.Dis. 1991, 143, 590; Suter et al. Am. Rev. Respir. Dis. 1992, 145,1016), acute pulmonary fibrotic diseases (Pan et al. Pathol. Int. 1996,46, 91), pulmonary sarcoidosis (Ishioka et al. Sarcoidosis VasculitisDiffuse Lung Dis. 1996, 13, 139), allergic respiratory diseases (Casaleet al. Am. J. Respir. Cell Mol. Biol. 1996, 15, 35), silicosis (Gossartet al. J. Immunol. 1996, 156, 1540; Vanhee et al. Eur. Respir. J. 1995,8, 834), coal worker's pneumoconiosis (Borm et al. Am. Rev. Respir. Dis.1988, 138, 1589), alveolar injury (Horinouchi et al. Am. J. Respir. CellMol. Biol. 1996, 14, 1044), hepatic failure (Gantner et al. J.Pharmacol. Exp. Therap. 1997, 280, 53), liver disease during acuteinflammation (Kim et al. J. Biol. Chem. 1997, 272, 1402), severealcoholic hepatitis (Bird et al. Ann. Intern. Med. 1990, 112, 917),malaria (Grau et al. Immunol. Rev. 1989, 112, 49; Taveme et al.Parasitol. Today 1996, 12, 290) including Plasmodium falciparum malaria(Perlmann et al. Infect. Immunit. 1997, 65, 116) and cerebral malaria(Rudin et al. Am. J. Pathol. 1997, 150, 257), non-insulin-dependentdiabetes mellitus (NIDDM; Stephens et al. J. Biol. Chem. 1997, 272, 971;Ofei et al. Diabetes 1996, 45, 881), congestive heart failure (Doyama etal. Int. J. Cardiol. 1996, 54, 217; McMurray et al. Br. Heart J. 1991,66, 356), damage following heart disease (Malkiel et al. Mol. Med. Today1996, 2, 336), atherosclerosis (Parums et al. J. Pathol. 1996, 179,A46), Alzheimer's disease (Fagarasan et al. Brain Res. 1996, 723, 231;Aisen et al. Gerontology 1997, 43, 143), acute encephalitis (Ichiyama etal. J. Neurol. 1996, 243, 457), brain injury (Cannon et al. Crit. CareMed. 1992, 20, 1414; Hansbrough et al. Surg. Clin. N. Am. 1987, 67, 69;Marano et al. Surg. Gynecol. Obstetr. 1990, 170, 32), multiple sclerosis(M. S.; Coyle. Adv. Neuroimmunol. 1996, 6, 143; Matusevicius et al. J.Neuroimmunol. 1996, 66, 115) including demyelation and oligiodendrocyteloss in multiple sclerosis (Brosnan et al. Brain Pathol. 1996, 6, 243),advanced cancer (MucWierzgon et al. J. Biol. Regulators HomeostaticAgents 1996, 10, 25), lymphoid malignancies (Levy et al. Crit. Rev.Immunol. 1996, 16, 31), pancreatitis (Exley et al. Gut 1992, 33, 1126)including systemic complications in acute pancreatitis (McKay et al. Br.J. Surg. 1996, 83, 919), impaired wound healing in infectioninflammation and cancer (Buck et al. Am. J. Pathol. 1996, 149, 195),myelodysplastic syndromes (Raza et al. Int. J. Hematol. 1996, 63, 265),systemic lupus erythematosus (Maury et al. Arthritis Rheum. 1989, 32,146), biliary cirrhosis (Miller et al. Am. J. Gasteroenterolog. 1992,87, 465), bowel necrosis (Sun et al. J. Clin. Invest. 1988, 81, 1328),psoriasis (Christophers. Austr. J. Dermatol. 1996, 37, S4), radiationinjury (Redlich et al. J. Immunol. 1996, 157, 1705), and toxicityfollowing administration of monoclonal antibodies such as OKT3 (Brod etal. Neurology 1996, 46, 1633). TNFα levels have also been related tohost-versus-graft reactions (Piguet et al. Immunol. Ser. 1992, 56, 409)including ischemia reperfusion injury (Colletti et al. J. Clin. Invest.1989, 85, 1333) and allograft rejections including those of the kidney(Maury et al. J. Exp. Med. 1987, 166, 1132), liver (Imagawa et al.Transplantation 1990, 50, 219), heart (Bolling et al. Transplantation1992, 53, 283), and skin (Stevens et al. Transplant. Proc. 1990, 22,1924), lung allograft rejection (Grossman et al. Immunol. Allergy Clin.N. Am. 1989, 9, 153) including chronic lung allograft rejection(obliterative bronchitis; LoCicero et al. J. Thorac. Cardiovasc. Surg.1990, 99, 1059), as well as complications due to total hip replacement(Cirino et al. Life Sci. 1996, 59, 86). TNFα has also been linked toinfectious diseases (review: Beutler et al. Crit. Care Med. 1993, 21,5423; Degre. Biotherapy 1996, 8, 219) including tuberculosis (Rook etal. Med. Malad. Infect. 1996, 26, 904), Helicobacter pylori infectionduring peptic ulcer disease (Beales et al. Gastroenterology 1997, 112,136), Chaga's disease resulting from Trypanosoma cruzi infection(Chandrasekar et al. Biochem. Biophys. Res. Commun. 1996, 223, 365),effects of Shiga-like toxin resulting from E. coli infection (Harel etal. J. Clin. Invest. 1992, 56, 40), the effects of enterotoxin Aresulting from Staphylococcus infection (Fischer et al. J. Immunol.1990, 144, 4663), meningococcal infection (Waage et al. Lancet 1987,355; Ossege et al. J. Neurolog. Sci. 1996, 144, 1), and infections fromBorrelia burgdorferi (Brandt et al. Infect. Immunol. 1990, 58, 983),Treponema pallidum (Chamberlin et al. Infect. Immunol. 1989, 57, 2872),cytomegalovirus (CMV; Geist et al. Am. J. Respir. Cell Mol. Biol. 1997,16, 31), influenza virus (Beutler et al. Clin. Res. 1986, 34, 491a),Sendai virus (Goldfield et al. Proc. Nat'l. Acad. Sci. USA 1989, 87,1490), Theiler's encephalomyelitis virus (Sierra et al. Immunology 1993,78, 399), and the human immunodeficiency virus (HIV; Poli. Proc. Nat'l.Acad. Sci. USA 1990, 87, 782; Vyakaram et al. AIDS 1990, 4, 21; Badleyet al. J. Exp. Med. 1997, 185, 55).

[0008] A number of diseases are thought to be mediated by excess orundesired matrix-destroying metalloprotease (MMP) activity or by animbalance in the ratio of the MMPs to the tissue inhibitors ofmetalloproteinases (TIMPs). These include osteoarthritis (Woessner etal. J. Biol. Chem. 1984, 259, 3633), rheumatoid arthritis (Mullins etal. Biochim. Biophys. Acta 1983, 695, 117; Woolley et al. ArthritisRheum. 1977, 20, 1231; Gravallese et al. Arthritis Rheum. 1991, 34,1076), septic arthritis (Williams et al. Arthritis Rheum. 1990, 33,533), tumor metastasis (Reich et al. Cancer Res. 1988, 48, 3307;Matrisian et al. Proc. Nat'l. Acad. Sci., USA 1986, 83, 9413),periodontal diseases (Overall et al. J. Periodontal Res. 1987, 22, 81),corneal ulceration (Burns et al. Invest. Opthalmol. Vis. Sci. 1989, 30,1569), proteinuria (Baricos et al. Biochem. J. 1988, 254, 609), coronarythrombosis from atherosclerotic plaque rupture (Henney et al. Proc.Nat'l. Acad. Sci., USA 1991, 88, 8154), aneurysmal aortic disease (Vineet al. Clin. Sci. 1991, 81, 233), birth control (Woessner et al.Steroids 1989, 54, 491), dystrophobic epidermolysis bullosa (Kronbergeret al. J. Invest. Dermatol. 1982, 79, 208), degenerative cartilage lossfollowing traumatic joint injury, osteopenias mediated by MMP activity,tempero mandibular joint disease, and demyelating diseases of thenervous system (Chantry et al. J. Neurochem. 1988, 50, 688).

[0009] Because inhibition of p38 leads to inhibition of TNFα productionand MMP production, inhibition of mitogen activated protein (MAP) kinasep38 enzyme provides an approach to the treatment of the above listeddiseases including osteoporosis and inflammatory disorders such asrheumatoid arthritis and COPD (Badger, A. M.; Bradbeer, J. N.; Votta,B.; Lee, J. C.; Adams, J. L.; Griswold, D. E. J. Pharm. Exper. Ther.1996, 279, 1453).

[0010] Vasculogenesis involves the de novo formation of blood vesselsfrom endothelial cell precursors or angioblasts. The first vascularstructures in the embryo are formed by vasculogenesis. Angiogenesisinvolves the development of capillaries from existing blood vessels, andis the principle mechanism by which organs, such as the brain and thekidney are vascularized. While vasculogenesis is restricted to embryonicdevelopment, angiogenesis can occur in the adult, for example duringpregnancy, the female cycle, or wound healing.

[0011] One major regulator of angiogenesis and vasculogenesis in bothembryonic development and some angiogenic-dependent diseases is vascularendothelial growth factor (VEGF; also called vascular permeabilityfactor, VPF). VEGF represents a family of isoforms of mitogens existingin homodimeric forms due to alternative RNA splicing. The VEGF isoformsare highly specific for vascular endothelial cells (for reviews, see:Farrara et al. Endocr. Rev. 1992, 13, 18; Neufield et al. FASEB J. 1999,13, 9). VEGF expression is induced by hypoxia (Shweiki et al. Nature1992, 359, 843), as well as by a variety of cytokines and growthfactors, such as interleukin-1, interleukin-6, epidermal growth factorand transforming growth factor.

[0012] To date, VEGF and the VEGF family members have been reported tobind to one or more of three transmembrane receptor tyrosine kinases(Mustonen et al. J. Cell Biol., 1995, 129, 895), VEGF receptor-1 (alsoknown as flt-i (fms-like tyrosine kinase-1)), VEGFR-2 (also known askinase insert domain containing receptor (KDR); the murine analogue ofKDR is known as fetal liver kinase-1 (flk-1)), and VEGFR-3 (also knownas flt-4). KDR and flt-1 have been shown to have different signaltransduction properties (Waltenberger et al. J. Biol. Chem. 1994, 269,26988); Park et al. Oncogene 1995, 10, 135). Thus, KDR undergoes strongligand-dependant tyrosine phosphorylation in intact cells, whereas flt-1displays a weak response. Thus, binding to KDR is a critical requirementfor induction of the full spectrum of VEGF-mediated biologicalresponses.

[0013] In vivo, VEGF plays a central role in vasculogenesis, and inducesangiogenesis and permeabilization of blood vessels. Deregulated VEGFexpression contributes to the development of a number of diseases thatare characterized by abnormal angiogenesis and/or hyperpermeabilityprocesses. Regulation of the VEGF-mediated signal transduction cascadewill therefore provide a useful mode for control of abnormalangiogenesis and/or hyperpermeability processes.

[0014] Angiogenesis is regarded as an absolute prerequisite for growthof tumors beyond about 1-2 mm. Oxygen and nutrients may be supplied tocells in tumor smaller than this limit through diffusion. However, everytumor is dependent on angiogenesis for continued growth after it hasreached a certain size. Tumorigenic cells within hypoxic regions oftumors respond by stimulation of VEGF production, which triggersactivation of quiescent endothelial cells to stimulate new blood vesselformation. (Shweiki et al. Proc. Nat'l. Acad. Sci., 1995, 92, 768). Inaddition, VEGF production in tumor regions where there is noangiogenesis may proceed through the ras signal transduction pathway(Grugel et al. J. Biol. Chem., 1995, 270, 25915; Rak et al. Cancer Res.1995, 55, 4575). In situ hybridization studies have demonstrated VEGFmRNA is strongly upregulated in a wide variety of human tumors,including lung (Mattern et al. Br. J. Cancer 1996, 73, 931), thyroid(Viglietto et al. Oncogene 1995, 11, 1569), breast (Brown et al. HumanPathol. 1995, 26, 86), gastrointestional tract (Brown et al. Cancer Res.1993, 53, 4727; Suzuki et al. Cancer Res. 1996, 56, 3004), kidney andbladder (Brown et al. Am. J. Pathol. 1993, 143I, 1255), ovary (Olson etal. Cancer Res. 1994, 54, 1255), and cervical (Guidi et al. J. Nat'lCancer Inst. 1995, 87, 12137) carcinomas, as well as angiosacroma(Hashimoto et al. Lab. Invest. 1995, 73, 859) and several intracranialtumors (Plate et al. Nature 1992, 359, 845; Phillips et al. Int. J.Oncol. 1993, 2, 913; Berkman et al. J. Clin. Invest., 1993, 91, 153).Neutralizing monoclonal antibodies to KDR have been shown to beefficacious in blocking tumor angiogenesis (Kim et al. Nature 1993, 362,841; Rockwell et al. Mol. Cell. Differ. 1995, 3, 315).

[0015] Overexpression of VEGF, for example under conditions of extremehypoxia, can lead to intraocular angiogenesis, resulting inhyperproliferation of blood vessels, leading eventually to blindness.Such a cascade of events has been observed for a number ofretinopathies, including diabetic retinopathy, ischemic retinal-veinocclusion, retinopathy of prematurity (Aiello et al. New Engl. J. Med.1994, 331, 1480; Peer et al. Lab. Invest. 1995, 72, 638), andage-related macular degeneration (AMD; see, Lopez et al. Invest.Opththalmol. Vis. Sci. 1996, 37, 855).

[0016] In rheumatoid arthritis (RA), the in-growth of vascular pannusmay be mediated by production of angiogenic factors. Levels ofimmunoreactive VEGF are high in the synovial fluid of RA patients, whileVEGF levels are low in the synovial fluid of patients with other formsof arthritis of with degenerative joint disease (Koch et al. J. Immunol.1994, 152, 4149). The angiogenesis inhibitor AGM-170 has been shown toprevent neovascularization of the joint in the rat collagen arthritismodel (Peacock et al. J. Exper. Med. 1992, 175, 1135).

[0017] Increased VEGF expression has also been shown in psoriatic skin,as well as bullous disorders associated with subepidermal blisterformation, such as bullous pemphigoid, erythema multiforme, anddermatitis herpetiformis (Brown et al. J. Invest. Dermatol. 1995, 104,744).

[0018] Because inhibition of KDR leads to inhibition of VEGF-mediatedangiogenesis and permeabilization, KDR inhibitors will be useful intreatment of diseases characterized by abnormal angiogenesis and/orhyperpermeability processes, including the above listed diseases.

SUMMARY OF THE INVENTION

[0019] The invention relates to a compound of formula (I)

[0020] wherein,

[0021] Y is OR¹ or NHR²,

[0022] Hal is chlorine or bromine,

[0023] R¹ is H or C₁-C₆ alkyl

[0024] R² is H, OH, CH₃ or CH₂OH,

[0025] Z¹ and Z² are each H or OH, wherein only one of Z¹ or Z² can beOH.

[0026] X¹ to X⁷ are each, independently, H, OH or O(CO)C₁-C₄ alkyl, and

[0027] n is 0 or 1,

[0028] with the proviso that at least one of conditions a-c is met,

[0029] a) Z¹ or Z² is OH,

[0030] b) R² is OH,

[0031] c) n is 1,

[0032] or a salt thereof, e.g., a pharmaceutically acceptable saltthereof, or an isolated stereoisomer thereof (collectively referred tohereinafter as the compounds of the invention). The term stereoisomer isunderstood to encompass diastereoisomers, enantiomers, geometricisomers, etc.

[0033] One of ordinary skill in the art will recognize that some of thecompounds of Formula (I) can exist in different geometrical isomericforms. In addition, some of the compounds of the present inventionpossess one or more asymmetric carbon atoms and are thus capable ofexisting in the form of optical isomers, as well as in the form ofracemic or nonracemic mixtures thereof, and in the form of diastereomersand diastereomeric mixtures. All of these compounds, including cisisomers, trans isomers, diastereomic mixtures, racemates, nonracemicmixtures of enantiomers, substantially pure, and pure enantiomers, areconsidered to be within the scope of the present invention. Herein,substantially pure enantiomers is intended to mean that no more than 5%w/w of the corresponding opposite enantiomer is present.

[0034] The optical isomers can be obtained by resolution of the racemicmixtures according to conventional processes, for example, by theformation of diastereoisomeric salts using an optically active acid orbase. Examples of appropriate acids are tartaric, diacetyltartaric,dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid. Mixturesof diastereoisomers can be separated into their individual diastereomerson the basis of their physical chemical differences by methods known tothose skilled in the art, for example, by chromatography or fractionalcrystallization. The optically active bases or acids are liberated fromthe separated diastereomeric salts. A different process for separationof optical isomers involves the use of a chiral chromatography column(e.g., chiral HPLC columns) optimally chosen to maximize the separationof the enantiomers. Suitable chiral HPLC columns are manufactured byDiacel, e.g., Chiracel OD and Chiracel OJ. The optically activecompounds of Formula (I) can likewise be obtained by utilizing opticallyactive starting materials.

[0035] The invention also comprises analogs of the compounds of theinvention.

[0036] Preference is given to compounds of the invention when n is 1.These compounds particularly include compounds of the invention whereinn is 1 in formula (I), Y is NHR² and R² is H or CH₃, compounds of theinvention wherein n is 1 in formula (I) and X¹ to X⁷ are each H,compounds of the invention wherein n is 1 in formula (I) and Z¹ and Z²are each H, compounds of the invention wherein n is 1 in formula (I) andZ¹ is H and Z² is OH or Z¹ is OH and Z² is H, compounds of the inventionwherein n is 1 in formula (I) and at least one of X¹ to X⁷ is OH orO(CO)C₁-C₄ alkyl, compounds of the invention wherein n is 1 in formula(I), Y is NHR² and R² is CH₂OH, compounds of the invention wherein n is1 in formula (I), Y is NHR² and R² is OH, and compounds of the inventionwherein n is 1 in formula (I) and Y is OH.

[0037] Other compounds of the invention of interest are those wherein informula (I) Z¹ is H and Z² is OH or Z¹ is OH and Z² is H. Theseparticularly include compounds of the invention wherein in formula (I)Z¹ is H and Z² is OH or Z¹ is OH and Z² is H, and n is 0, compounds ofthe invention wherein in formula (I) Z¹ is H and Z² is OH or Z¹ is OHand Z² is H, n is 0, Y is NHR² and R² is H or CH₃, compounds of theinvention wherein in formula (I) Z¹ is H and Z² is OH or Z¹ is OH and Z²is H, and n is 0 and X¹ to X⁷ are each H, compounds of the inventionwherein in formula (I) Z¹ is H and Z² is OH or Z¹ is OH and Z² is H, andn is 0 and at least one of X¹ to X⁷ is OH or O(CO)C₁-C₄ alkyl, compoundsof the invention wherein in formula (I) Z¹ is H and Z² is OH or Z¹ is OHand Z² is H, n is 0, Y is NHR² and R² is CH₂OH, compounds of theinvention wherein in formula (I) Z¹ is H and Z² is OH or Z¹ is OH and Z²is H, n is 0. Y is NHR² and R² is OH, and compounds of the inventionwherein in formula (1) Z¹ is H and Z² is OH or Z¹ is OH and Z² is H, andn is O and Y is OH.

[0038] Further compounds of the invention of interest are those whereinin formula (I), Y is NHR² and R² is OH. These compounds particularlyinclude compounds of the invention wherein in formula (I), Y is NHR²andR² is OH and n is 0, compounds of the invention wherein in formula (I),Y is NHR² and R² is OH and n is 0 and X¹ to X⁷ are each H, compounds ofthe invention wherein in formula (I), Y is NHR² and R² is OH and n is 0and Z¹ and Z² are each H, compounds of the invention wherein in formula(I), Y is NHR² and R² is OH and n is O and Z¹ is H and Z² is OH or Z¹ isOH and Z² is H, and compounds of the invention wherein in formula (I), Yis NHR² and R² is OH and n is 0 and at least one of X¹ to X⁷ is OH orO(CO)C₁-C₄ alkyl.

[0039] Compounds of the invention of interest are also those wherein informula (I) Y is OH. These compounds particularly include compounds ofthe invention wherein in formula (I) Y is OH and n is 0, compounds ofthe invention wherein in formula (I) Y is OH and n is 0 and X¹ to X⁷ areeach H, compounds of the invention wherein in formula (I) Y is OH and nis 0 and Z¹ and Z² are each H, compounds of the invention wherein informula (I) Y is OH and n is 0 and Z¹ is H and Z² is OH or Z¹ is OH andZ² is H, and compounds of the invention wherein in formula (I) Y is OHand n is 0 and at least one of X¹ to X⁷ is OH or O(CO)C₁-C₄ alkyl.

[0040] Particularly preferred compounds include:

[0041]4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide.

[0042]4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide.

[0043]4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide 1-oxide.

[0044]4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide 1-oxide.

[0045]4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-hydroxymethyl-2-pyridinecarboxamide 1-oxide.

[0046]4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-hydroxymethyl-2-pyridinecarboxamide 1-oxide, and salts, stereoisomers and prodrugs thereof.

[0047] A subgroup of the compounds of the invention which are ofinterest include compounds of formula (II), or a salt or stereoisomerthereof,

[0048] wherein,

[0049] Y is OR¹ or NHR²,

[0050] Hal is chlorine or bromine,

[0051] R¹ is H or C₁-C₆ alkyl

[0052] R² is H, OH, CH₃ or CH₂OH,

[0053] Z¹ and Z² are each H or OH, wherein only one of Z¹ or Z² can beOH,

[0054] X⁴ to X⁷ are each, independently, H, OH or O(CO)C₁-C₄ alkyl, and

[0055] n is 0 or 1,

[0056] with the proviso that at least one of conditions a-c is met,

[0057] a) Z¹ or Z² is OH,

[0058] b) R² is OH,

[0059] c) n is 1.

[0060] These include compounds of the invention wherein in formula (II)n is 1, compounds of the invention wherein in formula (II) n is 1 and Z¹and Z² are each H, compounds of the invention wherein in formula (II) nis 1, Z¹ and Z² are H and at least one of X⁴ to X⁷ is OH, compounds ofthe invention wherein in formula (II) n is 1, Z¹ and Z² are H and Y isNHR² and R² is H or CH₃, compounds of the invention wherein in formula(II) n is 0, compounds of the invention wherein in formula (II) n is 0and Z¹ is H and Z² is OH or Z¹ is OH and Z² is H, compounds of theinvention wherein in formula (II) n is 0, Z¹ and Z² are each H, and atleast one of X⁴ to X⁷ is OH, compounds of the invention wherein informula (II) n is 0 and Z¹ is H and Z² is OH or Z¹ is OH and Z² is H andat least one of X⁴ to X⁷ is OH, compounds of the invention wherein informula (II) n is 0 and Z¹ is H and Z² is OH or Z¹ is OH and Z² is H andY is NHR² and R² is H or CH₃, compounds of the invention wherein informula (II) n is 0 and Z¹ is H and Z² is OH or Z¹ is OH and Z² is H, Yis NHR^(¶)R² is OH, and compounds of the invention wherein in formula(II), Y is NHR², R² is OH, n is 0 and at least one of X⁴ to X⁷ is OH.

[0061] Another subgroup of the compounds of the invention which are ofinterest include compounds of formula (III), or a salt or isolatedstereoisomer thereof,

[0062] wherein,

[0063] Y is OR¹ or NHR²,

[0064] Hal is chlorine or bromine,

[0065] R¹ is H or C₁-C₆ alkyl

[0066] R² is H, OH, CH₃ or CH₂OH,

[0067] Z¹ and Z² are each H or OH, wherein only one of Z¹ or Z² can beOH, and

[0068] n is 0 or 1,

[0069] with the proviso that at least one of conditions a-c is met,

[0070] a) Z¹ or Z² is OH,

[0071] b) R² is OH,

[0072] c) n is 1.

[0073] These include compounds of the invention wherein in formula (III)n is 1 and Z¹ and Z² are each H, compounds of the invention wherein informula (III) n is 1, Z¹ and Z² are each H, Y is NHR² and R² is H orCH₃, compounds of the invention wherein in formula (III) n is 0 and Z¹is H and Z² is OH or Z¹ is OH and Z² is H, compounds of the inventionwherein in formula (III) n is 0, Z¹ is H and Z² is OH or Z¹ is OH and Z²is H, Y is NHR² and R² is H or CH₃, and compounds of the inventionwherein in formula (III) Y is OH.

[0074] The invention further relates to processes and methods ofpreparing the novel compounds of the invention. Such processes andmethods include, but are not limited to, the oxidation of the pyridylring of 4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridine carboxamide and4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide into their corresponding pyridine-1-oxides, the formaloxidation of any of the urea nitrogens of compounds of the inventioninto an N-hydroxyurea, the oxidation of any of the positions representedby X¹ to X⁷ of compounds of the invention whereby a hydrogen atom isreplaced by a hydroxyl group, the hydroxylation of the N-methyl amidesof4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide and4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide into the corresponding hydroxymethyl amides, thehydroxylation of said N-methyl amides into hydroxamic acids, thedemethylation of said N-methyl amides into unsubstituted amides, thehydrolysis of said N-methyl amides into carboxylic acids andcombinations thereof. Furthermore, the invention relates to theesterification of hydroxyl groups in the X¹ to X⁷ positions to, forexample, acetates.

[0075] Processes of interest include a process for preparing4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide, or pharmaceutically acceptable salt, or an isolatedstereoisomer thereof comprising oxidizing4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide into the corresponding pyridine-1-oxides and a process forpreparing4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or pharmaceutically acceptable salt, or an isolatedstereoisomer thereof comprising oxidizing4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide into the corresponding pyridine-1-oxides.

[0076] Compounds prepared by these methods are included in theinvention. Also included are compounds obtained by transformation,including metabolic transformation, of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide to either:

[0077] a) replace one or more of the phenyl hydrogens with a hydroxylgroup,

[0078] b) hydroxyate the N-methyl amide into a hydroxymethyl amide orhydroxamic acid,

[0079] c) demethylate the N-methyl amide into an unsubstituted amide,

[0080] d) oxidize one or more of the urea nitrogens from ═NH to ═NOH,

[0081] e) hydrolyze the N-methyl amide into a carboxylic acid,

[0082] f) oxidize the pyridine nitrogen into a pyridine-1-oxide, or

[0083] g) a combination of a-f,

[0084] with the proviso that at least one of steps b), d), and f) isperformed.

[0085] Of particular interest are compounds obtained by transformation,including metabolic transformation, of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridine carboxamide or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide to either:

[0086] a) hydroxyate the N-methyl amide into a hydroxymethyl amide orhydroxamic acid,

[0087] b) demethylate the N-methyl amide into an unsubstituted amide,

[0088] c) oxidize one or more of the urea nitrogens from ═NH to ═NOH,

[0089] d) hydrolyze the N-methyl amide into a carboxylic acid,

[0090] e) oxidize the pyridine nitrogen into a pyridine-1-oxide, or

[0091] f) a combination of a-e,

[0092] with the proviso that at least one of steps a), c), and e) isperformed.

[0093] It is understood that the term “pyridine-1-oxide” used throughoutthe document includes 1-oxo-pyridine and 1-hydroxy-pyridine, and thatfor the purposes of this document, all 3 terms are consideredinterchangeable. For example, ChemInnovation Software, Inc. Nomenclator™v. 3.01 identifies compounds of formula III where Y=NHCH₃, Hal=Cl, Z¹and Z⁷=H, and n=1, drawn in ChemDraw, asN-[4-chloro-3-(trifluoromethyl)phenyl]({4-[1-hydroxy-2-(N-methylcarbamoyl)(4-pyridyloxy)]phenyl}amino)carboxamide.

[0094] The invention further relates to a pharmaceutical compositioncomprising one or more compounds of the invention.

[0095] These include a pharmaceutical composition comprising aneffective amount of at least one compound of the invention and aphysiologically acceptable carrier. Preference is given to apharmaceutical composition comprising an effective amount of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or a pharmaceutically acceptable salt, an isolatedstereoisomer or a mixture thereof and a physiologically acceptablecarrier.

[0096] Pharmaceutically acceptable salts of these compounds are alsowithin the scope of the invention.

[0097] Salts of this invention are especially the pharmaceuticallyacceptable salts of compounds of formula (I) such as, for example,organic or inorganic acid addition salts of compounds of formula (I).Suitable inorganic acids include but are not limited to halogen acids(such as hydrochloric acid), sulfuric acid, or phosphoric acid. Suitableorganic acids include but are not limited to carboxylic, phosphonic,sulfonic, or sulfamic acids, with examples including acetic acid,propionic acid, octanoic acid, decanoic acid, dodecanoic acid, glycolicacid, lactic acid, 2- or 3-hydroxybutyric acid, γ-aminobutyric acid(GABA), gluconic acid, glucosemonocarboxylic acid, fumaric acid,succinic acid, adipic acid, pimelic acid, suberic acid, azeiaic acid,malic acid, tartaric acid, citric acid, glucaric acid, galactaric acid,amino acids (such as glutamic acid, aspartic acid, N-methylglycine,acetytaminoacetic acid, N-acetylasparagine or N-acetylcysteine), pyruvicacid, acetoacetic acid, methanesulfonic acid, 4-toluene sulfonic acid,benzenesulfonic acid, phosphoserine, and 2- or 3-glycerophosphoric acid.

[0098] Formation of prodrugs is well known in the art in order toenhance the properties of the parent compound; such properties includesolubility, absorption, biostability and release time (see“Pharmaceutical Dosage Form and Drug Delivery Systems” (Sixth Edition),edited by Ansel et al., published by Williams & Wilkins, pages 27-29,(1995) which is hereby incorporated by reference). Commonly usedprodrugs of the disclosed oxazolyl-phenyl-2,4-diamino-pyrimidinecompounds are designed to take advantage of the major drugbiotransformation reactions and are also to be considered within thescope of the invention. Major drug biotransformation reactions includeN-dealkylation, O-dealkylation, aliphatic hydroxylation, aromatichydroxylation, N-oxidation, S-oxidation, deamination, hydrolysisreactions, glucuronidation, sulfation and acetylation (see Goodman andGilman's The Pharmacological Basis of Therapeutics (Ninth Edition),editor Molinoff et al., publ. by McGraw-Hill, pages 11-13, (1996), whichis hereby incorporated by reference).

[0099] The invention also relates to methods for treating and preventingdiseases, for example, inflammatory and angiogenesis disorders andosteoporosis in mammals by administering a compound of the invention, ora pharmaceutical composition comprising a compound of the invention.

[0100] These include a method of treating or preventing osteoporosis,inflammation, and angiogenesis disorders (other than cancer) in a mammalby administering an effective amount of a compound of the invention tosaid mammal. Preference is given to a method of treating or preventingosteoporosis, inflammation, and angiogenesis disorders (other thancancer) in a mammal by administering an effective amount of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridine carboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or pharmaceutically acceptable salt, an isolatedstereoisomer or a mixture thereof to said mammal.

[0101] The invention also relates to a method of treating or preventingcancer and other hyperproliferative disorders by administering acompound of the invention, or a pharmaceutical composition comprisingone or more compounds of the invention, in combination with a cytotoxicagent.

[0102] These include a method of treating or preventing ahyper-proliferative disorder in a mammal by administering an effectiveamount of a compound of the invention to said mammal. Preference isgiven to a method of treating or preventing a hyper-proliferativedisorder in a mammal by administering an effective amount of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or a pharmaceutically acceptable salt, or anisolated stereoisomer or a mixture thereof to said mammal.

[0103] In the method of treating or preventing a hyper-proliferativedisorder in a mammal by administering an effective amount of a compoundsof the invention, one or more additional compounds or compositions maybe administered to said mammal, such as for example, an anticancercompound or composition, which is not a compound or compositionaccording to the invention, which is preferably a cytotoxic compound orcomposition. The method of treating or preventing a hyper-proliferativedisorder in a mammal also includes administering an effective amount of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or a pharmaceutically acceptable salt, or anisolated stereoisomer or a mixture thereof to said mammal together witha cytotoxic compound or composition.

[0104] Optional anti-proliferative agents which can be added to thecomposition include but are not limited to compounds listed on thecancer chemotherapy drug regimens in the 11^(th) Edition of the MerckIndex, (1996), which is hereby incorporated by reference, such asasparaginase, bleomycin, carboplatin, carmustine, chlorambucil,cisplatin, colaspase, cyclophosphamide, cytarabine, dacarbazine,dactinomycin, daunorubicin, doxorubicin (adriamycine), epirubicin,etoposide, 5-fluorouracil, hexamethylmelamine, hydroxyurea, ifosfamide,irinotecan, leucovorin, lomustine, mechlorethamine, 6-mercaptopurine,mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone,prednisone, procarbazine, raloxifen, streptozocin, tamoxifen,thioguanine, topotecan, vinblastine, vincristine, and vindesine.

[0105] Other anti-proliferative agents suitable for use with thecomposition of the invention include but are not limited to thosecompounds acknowledged to be used in the treatment of neoplasticdiseases in Goodman and Gilman 's The Pharmacological Basis ofTherapeutics (Ninth Edition), editor Molinoff et al., publ. byMcGraw-Hill, pages 1225-1287, (1996), which is hereby incorporated byreference such as aminoglutethimide, L-asparaginase, azathioprine,5-azacytidine cladribine, busulfan, diethylstilbestrol,2′,2′-difluorodeoxycytidine, docetaxel, erythrohydroxynonyladenine,ethinyl estradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridinemonophosphate, fludarabine phosphate, fluoxymesterone, flutamide,hydroxyprogesterone caproate, idarubicin, interferon,medroxyprogesterone acetate, megestrol acetate, melphalan, mitotane,paclitaxel, pentostatin, N-phosphonoacetyl-L-aspartate (PALA),plicamycin, semustine, teniposide, testosterone propionate, thiotepa,trimethylmelamine, uridine, and vinorelbine.

[0106] Other anti-proliferative agents suitable for use with thecomposition of the invention include but are not limited to otheranti-cancer agents such as oxaliplatin, gemcitabone, gefinitib,taxotere, BCNU, CCNU, DTIC, ara A, ara C, herceptin, actinomycin D,epothilone, irinotecan, raloxifen and topotecan.

[0107] Description of Treatment of Hyperproliferative Disorders

[0108] Cancer and hyperproliferative disorders are defined as follows.These disorders include but are not limited to solid tumors, such ascancers of the breast, respiratory tract, brain, reproductive organs,digestive tract, urinary tract, eye, liver, skin, head and neck,thyroid, parathyroid and their distant metastases. Those disorders alsoinclude lymphomas, sarcomas, and leukemias.

[0109] Examples of breast cancer include, but are not limited toinvasive ductal carcinoma, invasive lobular carcinoma, ductal carcinomain situ, and lobular carcinoma in situ.

[0110] Examples of cancers of the respiratory tract include, but are notlimited to small-cell and non-small-cell lung carcinoma, as well asbronchial adenoma and pleuropulmonary blastoma.

[0111] Examples of brain cancers include, but are not limited to brainstem and hypophtalmic glioma, cerebellar and cerebral astrocytoma,medulloblastoma, ependymoma, as well as neuroectodermal and pinealtumor.

[0112] Tumors of the male reproductive organs include, but are notlimited to prostate and testicular cancer.

[0113] Tumors of the female reproductive organs include, but are notlimited to endometrial, cervical, ovarian, vaginal, and vulvar cancer,as well as sarcoma of the uterus.

[0114] Tumors of the digestive tract include, but are not limited toanal, colon, colorectal, esophageal, gallblader, gastric, pancreatic,rectal, small-intestine, and salivary gland cancers.

[0115] Tumors of the urinary tract include, but are not limited tobladder, penile, kidney, renal pelvis, ureter, and urethral cancers.

[0116] Eye cancers include, but are not limited to intraocular melanomaand retinoblastoma.

[0117] Examples of liver cancers include, but are not limited tohepatocellular carcinoma (liver cell carcinomas with or withoutfibrolamellar variant), cholangiocarcinoma (intrahepatic bile ductcarcinoma), and mixed hepatocellular cholangiocarcinoma.

[0118] Skin cancers include, but are not limited to squamous cellcarcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skincancer, and non-melanoma skin cancer.

[0119] Head-and-neck cancers include, but are not limited tolaryngeal/hypopharyngeal/nasopharyngeal/oropharyngeal cancer, and lipand oral cavity cancer. Lymphomas include, but are not limited toAIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-celllymphoma, Hodgkin's disease, and lymphoma of the central nervous system.

[0120] Sarcomas include, but are not limited to sarcoma of the softtissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, andrhabdomyosarcoma. Leukemias include, but are not limited to acutemyeloid leukemia, acute lymphoblastic leukemia, chronic lymphocyticleukemia, chronic myelogenous leukemia, and hairy cell leukemia.

[0121] These disorders have been well characterized in man, but alsoexist with a similar etiology in other mammals, and can be treated bypharmaceutical compositions of the present invention.

[0122] Generally, the use of cytotoxic and/or cytostatic agents incombination with aryl urea compound raf kinase inhibitors will serve to(1) yield better efficacy in reducing the growth of a tumor or eveneliminate the tumor as compared to administration of either agent alone,(2) provide for the administration of lesser amounts of the administeredchemotherapeutic agents, (3) provide for a chemotherapeutic treatmentthat is well tolerated in the patient with fewer deleteriouspharmacological complications than observed with single agentchemotherapies and certain other combined therapies, (4) provide fortreating a broader spectrum of different cancer types in mammals,especially humans, (5) provide for a higher response rate among treatedpatients, (6) provide for a longer survival time among treated patientscompared to standard chemotherapy treatments, (7) provide a longer timefor tumor progression, and/or (8) yield efficacy and tolerabilityresults at least as good as those of the agents used alone, compared toknown instances where other cancer agent combinations produceantagonistic effects.

[0123] The present invention relates to a combination comprising (a) acompound according to the invention (b) at least one otherchemotherapeutic cytotoxic or cytostatic agent; or pharmaceuticallyacceptable salts of any component (a) or (b).

[0124] The invention also relates to a pharmaceutical preparation whichcomprises (1) quantities of (a) a compound according to the invention(b) at least one other cytotoxic or cytostatic agent in amounts whichare jointly effective for treating a cancer, where any component (a) or(b) can also be present in the form of a pharmaceutically acceptablesalt if at least one salt-forming group is present, with (2) one or morepharmaceutically acceptable carrier molecules.

[0125] The invention also relates to a method for treating a cancer thatcan be treated by administration of a compound according to theinvention and at least one other chemotherapeutic agent which is acytotoxic or cytostatic agent. The compound according to the inventionand the cytotoxic or cytostatic agent are administered to a mammal inquantities which together are therapeutically effective against hyperproliferative diseases as defined above. Thus, the compound according tothe invention is effective for raf kinase-mediated cancers. However,these compounds are also effective for cancers not mediated by rafkinase.

[0126] In a preferred embodiment, the present invention provides methodsfor treating a cancer in a mammal, especially a human patient,comprising administering an a compound according to the inventionoptionally in combination with a cytotoxic or cytostaticchemotherapeutic agent including but not limited to DNA topoisomerase Iand II inhibitors, DNA intercalators, alkylating agents, microtubuledisruptors, hormone and growth factor receptor agonists or antagonists,other kinase inhibitors and antimetabolites.

[0127] In another embodiment, a method is disclosed for administeringthe chemotherapeutic agents, including a compound according to theinvention and the cytotoxic and cytostatic agents, to the patient byoral delivery or by intravenous injection or infusion.

[0128] In another embodiment, the composition comprising a compoundaccording to the invention or the cytotoxic or cytostatic agent can beadministered to a patient in the form of a tablet, a liquid, a topicalgel, an inhaler or in the form of a sustained release composition.

[0129] In one embodiment of the invention, a compound according to theinvention can be administered simultaneously with a cytotoxic orcytostatic agent to a patient with a cancer, in the same formulation or,more typically in separate formulations and, often, using differentadministration routes. Administration can also be sequentially, in anyorder.

[0130] In another embodiment, a compound according to the invention canbe administered in tandem with the cytotoxic or cytostatic agent,wherein a compound according to the invention can be administered to apatient once or more per day for up to 28 consecutive days with theconcurrent or intermittent administration of a cytotoxic or cytostaticagent over the same total time period.

[0131] In another embodiment of the invention, a compound according tothe invention can be administered to a patient at an oral, intravenous,intramuscular, subcutaneous, or parenteral dosage which can range fromabout 0.1 to about 200 mg/kg of total body weight.

[0132] In another embodiment, the cytotoxic or cytostatic agent can beadministered to a patient at an intravenous, intramuscular,subcutaneous, or parenteral dosage which can range from about 0.1 mg to200 mg/kg of patient body weight.

[0133] Further, the invention relates to a method of inhibitingproliferation of cancer cells comprising contacting cancer cells with apharmaceutical preparation or product of the invention, especially amethod of treating a proliferative disease comprising contacting asubject, cells, tissues or a body fluid of said subject, suspected ofhaving a cancer with a pharmaceutical composition or product of thisinvention.

[0134] This invention also relates to compositions containing both acompound according to the invention and the other cytotoxic orcytostatic agents, in the amounts of this invention.

[0135] This invention further relates to kits comprising separate dosesof the two mentioned chemotherapeutic agents in separate containers. Thecombinations of the invention can also be formed in vivo, e.g., in apatient's body.

[0136] The term “cytotoxic” refers to an agent which can be administeredto kill or eliminate a cancer cell. The term “cytostatic” refers to anagent which can be administered to restrain tumor proliferation ratherthan induce cytotoxic cytoreduction yielding an elimination of thecancer cell from the total viable cell population of the patient. Thechemotherapeutic agents described herein, e.g., irinotecan, vinorelbine,gemcitabine, and paclitaxel are considered cytotoxic agents. Thesecytotoxic and cytostatic agents have gained wide spread use aschemotherapeutics in the treatment of various cancer types and are wellknown.

[0137] These and other cytotoxic/cytostatic agents can be administeredin the conventional formulations and regimens in which they are knownfor use alone.

General Preparative Methods

[0138] The compounds of the invention may be prepared by use of knownchemical reactions and procedures. Nevertheless, the following generalpreparative methods are presented to aid the reader in synthesizing thecompounds of the present invention, with more detailed particularexamples being presented below in the experimental section describingthe working examples.

[0139] All variable groups of these methods are as described in thegeneric description if they are not specifically defined below. When avariable group or substituent with a given symbol is used more than oncein a given structure, it is to be understood that each of these groupsor substituents may be independently varied within the range ofdefinitions for that symbol. It is recognized that compounds of theinvention with each claimed optional functional group cannot be preparedwith each of the below-listed methods. Within the scope of each methodoptional substituents are used which are stable to the reactionconditions, or the functional groups which may participate in thereactions are present in protected form where necessary, and the removalof such protective groups is completed at appropriate stages by methodswell known to those skilled in the art.

[0140] The compounds of the invention can be made according toconventional chemical methods, and/or as disclosed below, from startingmaterials which are either commercially available or producibleaccording to routine, conventional chemical methods. General methods forthe preparation of the compounds are given below, and the preparation ofrepresentative compounds is specifically illustrated in Examples 1 and2.

[0141] Ureas and hydroxyureas of formula (I) can be prepared by avariety of simple methods known in the art. General approaches for theformation of those compounds can be found in “Advanced OrganicChemistry”, by J. March, John Wiley and Sons, 1985 and in “ComprehensiveOrganic Transformations”, by R. C. Larock, VCH Publishers, 1989), whichare hereby incorporated by reference.

[0142] More specifically, the pyridine-1-oxides (n=1 in Formula (I)) ofthe present invention can be prepared from the corresponding pyridinesusing oxidation conditions known in the art. Some examples are asfollows:

[0143] peracids such as meta chloroperbenzoic acids in chlorinatedsolvents such as dichloromethane, dichloroethane, or chloroform(Markgraf et al., Tetrahedron 1991, 47, 183).

[0144] (Me₃SiO)₂ in the presence of a catalytic amount of perrhenic acidin chlorinated solvents such as dichloromethane (Coperet et al.,Tetrahedron Lett. 1998, 39, 761)

[0145] Perfluoro-cis-2-butyl-3-propyloxaziridine in several combinationsof halogenated solvents (Amone et al., Tetrahedron 1998, 54, 7831).

[0146] Hypofluoric acid—acetonitrile complex in chloroform (Dayan etal., Synthesis 1999, 1427).

[0147] Oxone, in the presence of a base such as KOH, in water (Robker etal., J. Chem. Res., Synop. 1993, 10, 412).

[0148] Magnesium monoperoxyphthalate, in the presence of glacial aceticacid (Klemm et al., J. Heterocyclic Chem. 1990, 6, 1537).

[0149] Hydrogen peroxide, in the presence of water and acetic acid (LinA. J., Org. Prep. Proced. Int. 1991, 23(1), 114).

[0150] Dimethyldioxirane in acetone (Boyd et al., J. Chem. Soc., PerkinTrans. 1991, 9, 2189).

[0151] The starting materials for the above-mentioned oxidations are bisaryl ureas, which contain a 2-acyl-pyridine in their side chains.Specific preparations of these ureas are already described in the patentliterature, and can be adapted to the compounds of the presentinvention. For example, Riedl, B., et al., “O-Carboxy Aryl SubstitutedDiphenyl Ureas as raf Kinase Inhibitors” PCT Int. Appl., WO 00 42012,Riedl, B., et al., “O-Carboxy Aryl Substituted Diphenyl Ureas as p3⁸Kinase Inhibitors” PCT Int. Appl., WO 00 41698.

[0152] Hydroxyureas of formula (I), where Z¹ is OH and Z² is H can beprepared as follows:

[0153] Substituted nitrobenzenes of Formula (II), which are known in theart, arc converted to hydroxyanilines of Formula (III), using a varietyof conditions known in the art, for example sodium borohydride in thepresence of transition metal catalysts (Yanada et al., Chem. Lett. 1989,951 and references cited therein), or N-methyldihydroacridine in thepresence of perchloric acid (Fukuzumi et al., J. Chem. Soc., PerkinTrans. II 1991, 9, 1393, and references cited therein).

[0154] In the second step, hydroxyanilines of Formula (III) can beconverted to the corresponding hydroxyureas by reaction with anisocyanate, or equivalent, in the same way ureas are being prepared.Examples of such reactions can be found in the art (Hoffman et al., J.Med. Chem. 1964, 7, 665, and Stoffel et al., Ber. Dtsch. Chem. Ges.1972, 105, 3115).

[0155] Similarly, hydroxyureas of formula (I), where Z¹ is H and Z² isOH can be prepared according to the same methods, by substituting thereagents in the appropriate way.

[0156] In both cases, the preparation of the arylamine fragment isillustrated in detail in the patent literature. For example, Miller S.et al, “Inhibition of p38 Kinase using Symmetrical and UnsymmetricalDiphenyl Ureas” PCT Int. Appl. WO 99 32463, Miller, S et al. “Inhibitionof raf Kinase using. Symmetrical and Unsymmetrical Substituted DiphenylUreas” PCT Int. Appl., WO 99 32436, Dumas, J. et al., “Inhibition of p38Kinase Activity using Substituted Heterocyclic Ureas” PCT Int. Appl., WO99 32111, Dumas, J. et al., “Method for the Treatment of Neoplasm byInhibition of raf Kinase using N-Heteroaryl-N′-(hetero)arylureas” PCTInt. Appl., WO 99 32106, Dumas, J. et al., “Inhibition of p38 KinaseActivity using Aryl- and Heteroaryl-Substituted Heterocyclic Ureas” PCTInt. Appl., WO 99 32110, Dumas, J., et al., “Inhibition of raf Kinaseusing Aryl- and Heteroaryl-Substituted Heterocyclic Ureas” PCT Int.Appl., WO 99 32455, Riedl, B., et al., “O-Carboxy Aryl SubstitutedDiphenyl Ureas as raf Kinase Inhibitors” PCT Int. Appl., WO 00 42012,Riedl, B., et al., “O-Carboxy Aryl Substituted Diphenyl Ureas as p38Kinase Inhibitors” PCT Int. Appl., WO 00 41698.

[0157] Hydroxymethyl amides of Formula (I) where Y is NHCH₂OH can beprepared by hydroxylation of the corresponding unsubstituted amides(Y=NH₂) by a variety of methods known in the art, for example aqueousformaldehyde in the presence of ethanol and sodium hydroxide (Weaver etal., J. Org. Chem. 1951, 16, 1111), or in the presence of potassiumcarbonate (Haworth et al., J. Chem. Soc. 1946, 1003).

[0158] Hydroxamic acids of Formula (I) where Y is NHOH can be preparedby amidation of the corresponding esters (Y=O alkyl) by a variety ofmethods known in the art, for example hydroxylamine in the presence ofacetic acid and water (Boshagen, H., Ber. Dtsch. Chem. Ges. 1967, 100,954). The same compounds can be obtained from the corresponding acids(Y=OH) by one pot activation of the acid with ethyl chloroformate,followed by reaction with hydroxylamine in methanol (Reddy et al.,Tetrahedron Lett. 2000, 41(33), 6285), or by activation of the acid intoan 1-acylimidazole, followed by reaction with hydroxylaminehydrochloride (Staab et al., Angewandte Chem., 1962, 74, 407).

[0159] Finally, ureas may be further manipulated using methods familiarto those skilled in the art.

[0160] The invention also includes pharmaceutical compositions includinga compound of the invention, and a physiologically acceptable carrier.

[0161] The compounds may be administered orally, topically,parenterally, by injection, by inhalation or spray or rectally in dosageunit formulations. Administration by injection includes intravenous,intramuscular, subcutaneous and parenteral injections, as well as use ofinfusion techniques. One or more compounds may be present in associationwith one or more non-toxic pharmaceutically acceptable carriers and ifdesired other active ingredients.

[0162] Compositions intended for oral use may be prepared according toany suitable method known to the art for the manufacture ofpharmaceutical compositions. Such compositions may contain one or moreagents selected from the group consisting of diluents, sweeteningagents, flavoring agents, coloring agents and preserving agents in orderto provide palatable preparations. Tablets contain the active ingredientin admixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be,for example, inert diluents, such as calcium carbonate, sodiumcarbonate, lactose, calcium phosphate or sodium phosphate; granulatingand disintegrating agents, for example, corn starch, or alginic acid;and binding agents, for example magnesium stearate, stearic acid ortalc. The tablets may be uncoated or they may be coated by knowntechniques to delay disintegration and adsorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. These compounds mayalso be prepared in solid, rapidly released form.

[0163] Formulations for oral use may also be presented as hard gelatincapsules wherein the active ingredient is mixed with an inert soliddiluent, for example, calcium carbonate, calcium phosphate or kaolin, oras soft gelatin capsules wherein the active ingredient is mixed withwater or an oil medium, for example peanut oil, liquid paraffin or oliveoil.

[0164] Aqueous suspensions contain the active materials in admixturewith excipients suitable for the manufacture of aqueous suspensions.Such excipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally occurring phosphatide,for example, lecithin, or condensation products or an alkylene oxidewith fatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolsuch as polyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives, for exampleethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, oneor more flavoring agents, and one or more sweetening agents, such assucrose or saccharin.

[0165] Dispersible powders and granules suitable for preparation of anaqueous suspension by the addition of water provide the activeingredient in admixture with a dispersing or wetting agent, suspendingagent and one or more preservatives. Suitable dispersing or wettingagents and suspending agents are exemplified by those already mentionedabove. Additional excipients, for example, sweetening, flavoring andcoloring agents, may also be present.

[0166] The compounds may also be in the form of non-aqueous liquidformulations, e.g., oily suspensions which may be formulated bysuspending the active ingredients in a vegetable oil, for examplearachis oil, olive oil, sesame oil or peanut oil, or in a mineral oilsuch as liquid paraffin. The oily suspensions may contain a thickeningagent, for example beeswax, hard paraffin or cetyl alcohol. Sweeteningagents such as those set forth above, and flavoring agents may be addedto provide palatable oral preparations. These compositions may bepreserved by the addition of an anti-oxidant such as ascorbic acid.

[0167] Pharmaceutical compositions of the invention may also be in theform of oil-in-water emulsions. The oily phase may be a vegetable oil,for example olive oil or arachis oil, or a mineral oil, for exampleliquid paraffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

[0168] Syrups and elixirs may be formulated with sweetening agents, forexample glycerol, propylene glycol, sorbitol or sucrose. Suchformulations may also contain a demulcent, a preservative and flavoringand coloring agents.

[0169] The compounds may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include cocoa butter and polyethylene glycols.

[0170] For all regimens of use disclosed herein for compounds of theinvention, the daily oral dosage regimen will preferably be from 0.01 to200 mg/kg of total body weight. The daily dosage for administration byinjection, including intravenous, intramuscular, subcutaneous andparenteral injections, and use of infusion techniques will preferably befrom 0.01 to 200 mg/kg of total body weight. The daily rectal dosageregime will preferably be from 0.01 to 200 mg/kg of total body weight.The daily topical dosage regime will preferably be from 0.1 to 200 mgadministered between one to four times daily. The daily inhalationdosage regime will preferably be from 0.01 to 10 mg/kg of total bodyweight. The dosage units employed to provide these dosage regimes can beadministered on a daily basis, one or more times, or for extendedperiods, such as on a weekly or monthly basis.

[0171] It will be appreciated by those skilled in the art that theparticular method of administration will depend on a variety of factors,all of which are considered routinely when administering therapeutics.It will also be appreciated by one skilled in the art that the specificdose level for a given patient depends on a variety of factors,including specific activity of the compound administered, age, bodyweight, health, sex, diet, time and route of administration, rate ofexcretion, etc. It will be further appreciated by one skilled in the artthat the optimal course of treatment, i.e., the mode of treatment andthe daily number of doses of a compound of the invention for a definednumber of days, can be ascertained by those skilled in the art usingconventional treatment tests.

[0172] The compounds can be produced from known compounds (or fromstarting materials which, in turn, can be produced from knowncompounds), e.g., through the general preparative methods disclosedherein. The activity of a given compound to inhibit raf, p38, or KDR(VEGFR²) kinases can be routinely assayed, e.g., according to proceduresdisclosed herein.

[0173] The entire enclosure of all applications, patents andpublications cited above and below are hereby incorporated by reference,including non-provisional application Ser. No. 09/425,228 filed Oct. 22,1999, and non-provisional application Ser. No. 09/458,548 filed Jan. 12,2001.

[0174] Without further elaboration, it is believed that one skilled inthe art can, using the preceding description, utilize the presentinvention to its fullest extent. The following examples are, therefore,to be construed as merely illustrative and not limitative of theremainder of the disclosure in any way whatsoever.

EXAMPLES

[0175] All reactions were performed in flame-dried or oven-driedglassware under a positive pressure of dry argon or dry nitrogen, andwere stirred magnetically unless otherwise indicated. Sensitive liquidsand solutions were transferred via syringe or cannula, and introducedinto reaction vessels through rubber septa. Unless otherwise stated, theterm ‘concentration under reduced pressure’ refers to use of a Buchirotary evaporator at approximately 15 mmHg. Unless otherwise stated, theterm ‘under high vacuum’ refers to a vacuum of 0.4-1.0 mmHg.

[0176] All temperatures are reported uncorrected in degrees Celsius (°C.). Unless otherwise indicated, all parts and percentages are byweight. Commercial grade reagents and solvents were used without furtherpurification.

[0177] Thin-layer chromatography (TLC) is performed using Whatman®pre-coated glass-backed silica gel 60A F-254 250 μm plates.Visualization of plates is effected by one or more of the followingtechniques: (a) ultraviolet illumination, (b) exposure to iodine vapor,(c) immersion of the plate in a 10% solution of phosphomolybdic acid inethanol followed by heating, (d) immersion of the plate in a ceriumsulfate solution followed by heating, and/or (e) immersion of the platein an acidic ethanol solution of 2,4-dinitrophenylhydrazine followed byheating. Column chromatography (flash chromatography) is performed using230-400 mesh EM Science® silica gel.

[0178] Melting points (mp) are determined using a Thomas-Hoover meltingpoint apparatus or a Mettler FP66 automated melting point apparatus andare uncorrected. Fourier transform infrared spectra are obtained using aMattson 4020 Galaxy Series spectrophotometer. Proton (¹H) nuclearmagnetic resonance (NMR) spectra are measured with a General ElectricGN-Omega 300 (300 MHz) spectrometer with either Me₄Si (δ 0.00) orresidual protonated solvent (CHCl₃ δ 7.26; MeOH δ 3.30; DMSO δ 2.49) asstandard. Carbon (¹³C) NMR spectra are measured with a General ElectricGN-Omega 300 (75 MHz) spectrometer with solvent (CDCl₃ δ 77.0; MeOD-d₃;δ 49.0; DMSO-d₆ δ 39.5) as standard. Low resolution mass spectra (MS)and high resolution mass spectra (HRMS) are either obtained as electronimpact (EI) mass spectra or as fast atom bombardment (FAB) mass spectra.Electron impact mass spectra (EI-MS) are obtained with a Hewlett Packard5989A mass spectrometer equipped with a Vacumetrics Desorption ChemicalIonization Probe for sample introduction. The ion source is maintainedat 250° C. Electron impact ionization is performed with electron energyof 70 eV and a trap current of 300 μA. Liquid-cesium secondary ion massspectra (FAB-MS), an updated version of fast atom bombardment areobtained using a Kratos Concept 1-H spectrometer. Chemical ionizationmass spectra (CI-MS) are obtained using a Hewlett Packard MS-Engine(5989A) with methane or ammonia as the reagent gas (1×10⁴ torr to2.5×10⁻⁴ torr). The direct insertion desorption chemical ionization(DCI) probe (Vaccumetrics, Inc.) is ramped from 0-1.5 amps in 10 sec andheld at 10 amps until all traces of the sample disappeared (˜1-2 min).Spectra are scanned from 50-800 amu at 2 sec per scan. HPLC—electrospraymass spectra (HPLC ES-MS) are obtained using a Hewlett-Packard 1100 HPLCequipped with a quaternary pump, a variable wavelength detector, a C-18column, and a Finnigan LCQ ion trap mass spectrometer with electrosprayionization. Spectra are scanned from 120-800 amu using a variable iontime according to the number of ions in the source. Gaschromatography—ion selective mass spectra (GC-MS) are obtained with aHewlett Packard 5890 gas chromatograph equipped with an HP-1 methylsilicone column (0.33 mM coating; 25 m—0.2 mm) and a Hewlett Packard5971 Mass Selective Detector (ionization energy 70 eV). Elementalanalyses are conducted by Robertson Microlit Labs, Madison N.J.

[0179] All compounds displayed NMR spectra, LRMS and either elementalanalysis or HRMS consistent with assigned structures.

Example 1 Preparation ofN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)-1-oxo-(4-pyridyloxy)]phenyl}urea

[0180]

[0181] To a stirred mixture ofN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)(4-pyridyloxy)]phenyl}urea(500 mg, 1.08 mmol), in a mixture of anh CH₂Cl₂ (2.2 mL) and anh THF(2.2 mL) was added 3-chloroperbenzoic acid (77% pure, 1.09 g, 4.86 mmol,4.5 equiv.), and the resulting mixture was heated at 40° C. for 33 h.The resulting mixture was concentrated under reduced pressure, and thecrude product was purified by MPLC (Biotage®; gradient from 20%acetone/hexane to 50% acetone/hexane). Recrystallization from EtOAcaffordedN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)-1-oxo-(4-pyridyloxy)]phenyl}ureaas a white solid (293 mg, 57%): mp (uncorrected) 232-234° C.; TLC (50%acetone/hexane) R_(f) 0.13; ¹H-NMR (DMSO-d₆) δ 11.48 (broad s, 1H), 9.19(s, 1H), 8.98 (s, 1H), 8.38 (d, J=5.8 Hz, 1H), 8.10 (d, J=2.5 Hz, 1H),7.64 (dd, J=8.2 Hz, 2.6 Hz, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.57 (d, J=8.7Hz, 2H), 7.54 (d, J=2.6 Hz, 1H), 7.28 (dd, J=5.7 Hz, 2.5 Hz, 1H), 7.18(d, J=8.8 Hz, 2H), 2.86 (d, J=5.0 Hz, 3H); HPLC EI-MS m/z 481 ((M+H)⁺).Anal. calcd for C₂₁H₁₆ClFN₄O₄: C, 52.46%; H, 3.33%; N, 11.65%. Found: C,52.22%; H, 3.39%; N, 11.49%.

Example 2 Preparation ofN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-1-oxo-(4-pyridyloxy)]phenyl}urea

[0182] Step 1: Preparation of 4-chloro-2-pyridinecarboxamide

[0183] To a stirred mixture of methyl 4-chloro-2-pyridinecarboxylatehydrochloride (1.0 g, 4.81 mmol) dissolved in conc. aqueous ammonia (32mL) was added ammonium chloride (96.2 mg, 1.8 mmol, 0.37 equiv.), andthe heterogeneous reaction mixture was stirred at ambient temperaturefor 16 h. The reaction mixture was poured into EtOAc (500 mL) and water(300 mL). The organic layer was washed with water (2×300 mL) and asaturated NaCl solution (1×300 mL), dried (MgSO₄), concentrated in vacuoto give 4-chloro-2-pyridinecarboxamide as a beige solid (604.3 mg,80.3%): TLC (50% EtOAc/hexane) R_(f) 0.20; ¹H-NMR (DMSO-d₆) δ 8.61 (d,J=5.4 Hz, 1H), 8.20 (broad s, 1H), 8.02 (d, J=1.8 Hz, 1H), 7.81 (broads, 1H), 7.76 to 7.73 (m, 1H).

[0184] Step 2: Preparation of 4-(4-aminophenoxy)-2-pyridinecarboxamide

[0185] To 4-aminophenol (418 mg, 3.83 mmol) in anh DMF(7.7 mL) was addedpotassium tert-butoxide (447 mg, 3.98 mmol, 1.04 equiv.) in one portion.The reaction mixture was stirred at room temperature for 2 h, and asolution of 4-chloro-2-pyridinecarboxamide (600 mg, 3.83 mmol, 1.0equiv.) in anh DMF (4 mL) was then added. The reaction mixture wasstirred at 80° C. for 3 days and poured into a mixture of EtOAc and asaturated NaCl solution. The organic layer was sequentially washed witha saturated NH₄Cl solution then a saturated NaCl solution, dried(MgSO₄), and concentrated under reduced pressure. The crude product waspurified using MPLC chromatography (Biotage®; gradient from 100% EtOActo followed by 10% MeOH/50% EtOAc/40% hexane) to give the4-chloro-5-trifluoromethylaniline as a brown solid (510 mg, 58%). ¹H-NMR(DMSO-d₆) δ 8.43 (d, J=5.7 Hz, 1H), 8.07 (br s, 1H), 7.66 (br s, 1H),7.31 (d, J=2.7 Hz, 1H), 7.07 (dd, J=5.7 Hz, 2.7 Hz, 1H), 6.85 (d, J=9.0Hz, 2 H), 6.62 (d, J=8.7 Hz, 2H), 5.17 (broad s, 2H); HPLC EI-MS m/z 230((M+H)⁺.

[0186] Step 3: Preparation ofN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-(4-pyridyloxy)]phenyl}urea

[0187] A mixture of 4-chloro-5-trifluoromethylaniline (451 mg, 2.31mmol, 1.1 equiv.) and 1,1′-carbonyl diimidazole (419 mg, 2.54 mmol, 1.2equiv.) in anh dichloroethane (5.5 mL) was stirred under argon at 65° C.for 16 h. Once cooled to room temperature, a solution of4-(4-aminophenoxy)-2-pyridinecarboxamide (480 mg, 2.09 mmol) in anh THF(4.0 mL) was added, and the reaction mixture was stirred at 60° C. for 4h. The reaction mixture was poured into EtOAc, and the organic layer waswashed with water (2×) and a saturated NaCl solution (1×), dried(MgSO₄), filtered, and evaporated in vacuo. Purification using MPLCchromatography (Biotage®; gradient from 100% EtOAc to 2% MeOH/EtOAc)gaveN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-(4-pyridyloxy)]phenyl}ureaas a white solid (770 mg, 82%): TLC (EtOAc) R_(f) 0.11, 100% ethylacetate ¹H-NMR (DMSO-d₆) δ 9.21 (s, 1H), 8.99 (s, 1H), 8.50 (d, J=5.6Hz, 1H), 8.11 (s, 1H), 8.10 (s, 1H), 7.69 (broad s, 1H), 7.64 (dd, J=8.2Hz, 2.1 Hz, 1H), 7.61 (s, 1H), 7.59 (d, J=8.8 Hz, 2H), 7.39 (d, J=2.5Hz, 1H), 7.15 (d, J=8.9 Hz, 2H), 7.14 (m, 1H); MS LC-MS (MH+=451). Anal.calcd for C₂₀H₁₄ClF₃N₄O₃: C, 53.29%; H, 3.13%; N, 12.43%. Found: C,53.33%; H, 3.21%; N, 12.60%.

[0188] Step 4: Preparation ofN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-1-oxo-(4-pyridyloxy)]phenyl}urea

[0189]N-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-1-oxo-(4-pyridyloxy)]phenyl}urea(125.6 mg, 51%) was prepared as a white solid fromN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-(4-pyridyloxy)]phenyl}urea(240.0 mg, 0.53 mmol), in the manner described forN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)-1-oxo-(4-pyridyloxy)]phenyl}urea:TLC (5% MeOH/CH₂Cl₂) R_(f) 0.17; ¹H-NMR (DMSO-d₆) δ 10.72 (d, J=4.3 Hz,1H), 9.21 (s, 1H), 8.99 (s, 1H), 8.36 (d, J=7.2 Hz, 1H), 8.31 (d, J=4.1Hz, 1H), 8.10 (d, J=2.3 Hz, 1H), 7.65 (dd, J=8.7 Hz, 2.3 Hz, 1H), 7.60(d, J=8.9 Hz, 1H), 7.57 (d, J=9.0 Hz, 2H), 7.54 (d, J=3.8 Hz, 1H), 7.28(dd, J=7.2 Hz, 3.8 Hz, 1H), 7.18 (d, J=9.0 Hz, 2 H); HPLC EI-MS m/z 467((M+H)⁺; Anal. calcd for C₂₀H₁₄ClF₃N₄O₄ 0.5H₂O: C, 50.49%; H, 3.18%; N,11.78%. Found. C, 50.69%; H, 2.86%; N, 11.47%.

BIOLOGICAL EXAMPLES

[0190] P38 Kinase in vitro Assay

[0191] The in vitro inhibitory properties of compounds were determinedusing a p38 kinase inhibition assay. P38 activity was detected using anin vitro kinase assay run in 96-well microtiter plates. Recombinanthuman p38 (0.5 μg/mL) was mixed with substrate (myelin basic protein, 5μg/mL) in kinase buffer (25 mM Hepes, 20 mM MgCl₂ and 150 mM NaCl) andcompound. One μCi/well of ³³P-labeled ATP (10 μM) was added to a finalvolume of 100 μL. The reaction was run at 32° C. for 30 min. and stoppedwith a 1M HCl solution. The amount of radioactivity incorporated intothe substrate was determined by trapping the labeled substrate ontonegatively charged glass fiber filter paper using a 1% phosphoric acidsolution and read with a scintillation counter. Negative controlsinclude substrate plus ATP alone.

[0192] LPS Induced TNFα Production in Mice

[0193] The in vivo inhibitory properties of selected compounds can bedetermined using a murine LPS induced TNFα production in vivo model.BALB/c mice (Charles River Breeding Laboratories; Kingston, N.Y.) ingroups of ten were treated with either vehicle or compound by the routenoted. After one hour, endotoxin (E. coli lipopolysaccharide (LPS) 100μg) was administered intraperitoneally (i.p.). After 90 min, animalswere euthanized by carbon dioxide asphyxiation and plasma was obtainedfrom individual animals by cardiac puncture into heparinized tubes. Thesamples were clarified by centrifugation at 12,500×g for 5 min at 4° C.The supernatants were decanted to new tubes, which were stored as neededat −20° C. TNFα levels in sera were measured using a commercial murineTNF ELISA kit (Genzyme).

[0194] The two preceding biological examples can be used to demonstratethat the compounds are inhibiting p38 kinase in vitro and in vivo, andtherefore establishes their utility in the treatment of p38 mediateddiseases, such as inflammation and osteoporosis.

[0195] In Vitro raf Kinase Assay

[0196] In an in vitro kinase assay, raf was incubated with MEK in 20 mMTris-HCl, pH 8.2 containing 2 mM 2-mercaptoethanol and 100 mM NaCl. Thisprotein solution (20 μL) was mixed with water (5 μL) or with compoundsdiluted with distilled water from 10 mM stock solutions of compoundsdissolved in DMSO. The kinase reaction was initiated by adding 25 μL[γ-³³P]ATP (1000-3000 dpm/pmol) in 80 mM Tris-HCl, pH 7.5, 120 mM NaCl,1.6 mM DTT, 16 mM MgCl₂. The reaction mixtures were incubated at 32° C.,usually for 22 min. Incorporation of ³³P into protein was assayed byharvesting the reaction onto phosphocellulose mats, washing away freecounts with a 1% phosphoric acid solution and quantitatingphosphorylation by liquid scintillation counting. For high throughputscreening, 10 μM ATP and 0.4 μM MEK are used. In some experiments, thekinase reaction is stopped by adding an equal amount of Laemmli samplebuffer. Samples are boiled 3 min and the proteins resolved byelectrophoresis on 7.5% Laemmli gels. Gels were fixed, dried and exposedto an imaging plate (Fuji). Phosphorylation was analyzed using a FujixBio-Imaging Analyzer System. Compounds of Examples 1 and 2 show>50%inhibition at 10 micromolars in this assay, which is a marked inhibitionof raf kinase in vitro.

[0197] Tumor Cell Proliferation Assay

[0198] For in vitro growth assay, human tumor cell lines, including butnot limited to HCT 116 and DLD-1, containing mutated K-ras genes wereused in standard proliferation assays for anchorage dependent growth onplastic or anchorage independent growth in soft agar. Human tumor celllines were obtained from ATCC (Rockville Md.) and maintained in RPMIwith 10% heat inactivated fetal bovine serum and 200 mM glutamine. Cellculture media and additives were obtained from Gibco/BRL (Gaithersburg,Md.) except for fetal bovine serum (JRH Biosciences, Lenexa, Kans.). Ina standard proliferation assay for anchorage dependent growth, 3×10³cells were seeded into 96-well tissue culture plates and allowed toattach overnight at 37° C. in a 5% CO₂ incubator. Compounds weretitrated in media in dilution series and added to 96 well cell cultures.Cells were allowed to grow 5 days typically with a feeding of freshcompound containing media on day three. Proliferation was monitored bymeasuring metabolic activity with standard XTT colorimetric assay(Boehringer Mannheim) measured by standard ELISA plate reader at OD490/560, harvesting the cells onto glass fiber mats using a cellharvester and measuring ³H-thymidine incorporation by liquid scintillantcounting.

[0199] For anchorage independent cell growth, cells were plated at 1×10³to 3×10³ in 0.4% Seaplaque agarose in RPMI complete media, overlaying abottom layer containing only 0.64% agar in RPMI complete media in24-well tissue culture plates. Complete media plus dilution series ofcompounds were added to wells and incubated at 37° C. in a 5% CO₂incubator for 10-14 days with repeated feedings of fresh mediacontaining compound at 3-4 day intervals. Colony formation was monitoredand total cell mass, average colony size and number of colonies werequantitated using image capture technology and image analysis software(Image Pro Plus, media Cybernetics).

[0200] The two preceding assays establish that the compounds of FormulaI are active to inhibit raf kinase activity and to inhibit oncogeniccell growth.

[0201] KDR (VEGFR2) Assay

[0202] The cytosolic kinase domain of KDR kinase is expressed as a 6Hisfusion protein in Sf9 insect cells. The KDR kinase domain fusion proteinis purified over a Ni++ chelating column. Ninety-six well ELISA platesare coated with 5 μg poly(Glu4; Tyr1) (Sigma Chemical Co., St Louis,Mo.) in 100 μl HEPES buffer (20 mM HEPES, pH 7.5, 150 mM NaCl, 0.02%Thimerosal) at 4° overnight. Before use, the plate is washed with HEPES,NaCl buffer and the plates are blocked with 1% BSA, 0.1% Tween 20 inHEPES, NaCl buffer.

[0203] Test compounds are serially diluted in 100% DMSO from 4 mM to0.12 μM in half-log dilutions. These dilutions are further dilutedtwenty fold in H₂O to obtain compound solutions in 5% DMSO. Followingloading of the assay plate with 85 μl of assay buffer (20 mM HEPES, pH7.5, 100 mM KCl, 10 mM MgCl₂, 3 mM MnCl₂, 0.05% glycerol, 0.005% TritonX-100, 1 mM -mercaptoethanol, with or without 3.3 μM ATP), 5 μl of thediluted compounds are added to a final assay volume of 100 μl. Finalconcentrations are between 10 μM, and 0.3 nM in 0.25% DMSO. The assay isinitiated by the addition of 10 μl (30 ng) of KDR kinase domain.

[0204] The assay is incubated with test compound or vehicle alone withgentle agitation at room temperature for 60 minutes. The wells arewashed and phosphotyrosines (PY) are probed with an anti-phosphotyrosine(PY), mAb clone 4G10 (Upstate Biotechnology, Lake Placid, N.Y.).PY/anti-PY complexes are detected with an anti-mouse IgG/HRP conjugate(Amersham International plc, Buckinghamshire, England). Phosphotyrosineis quantitated by incubating with 100 μl 3,3′,5,5′ tetramethylbenzidinesolution (Kirkegaard and Perry, TMB Microwell 1 Component peroxidasesubstrate). Color development is arrested by the addition of 100 μl 1%HCl-based stop solution (Kirkegaard and Perry, TMB 1 Component StopSolution).

[0205] Optical densities are determined spectrophotometrically at 450 nmin a 96-well plate reader, SpectraMax 250 (Molecular Devices).Background (no ATP in assay) OD values are subtracted from all ODs andthe percent inhibition is calculated according to the equation:${\% \quad {Inhibition}} = \frac{( {{{OD}( {{vehicle}\quad {control}} )} - {{OD}( {{with}\quad {compound}} )}} ) \times 100}{{{OD}( {{vehicle}\quad {control}} )} - {{OD}( {{no}\quad {ATP}\quad {added}} )}}$

[0206] The IC₅₀ values are determined with a least squares analysisprogram using compound concentration versus percent inhibition.

[0207] Cell Mechanistic Assay-Inhibition of 3T3 KDR Phosphorylation

[0208] NIH3T3 cells expressing the full length KDR receptor are grown inDMEM (Life Technologies, Inc., Grand Island, N.Y.) supplemented with 10%newborn calf serum, low glucose, 25 mM /L sodium pyruvate, pyridoxinehydrochloride and 0.2 mg/ml of G418 (Life Technologies Inc., GrandIsland, N.Y.). The cells are maintained in collagen I-coated T75 flasks(Becton Dickinson Labware, Bedford, Mass.) in a humidified 5% CO2atmosphere at 37° C.

[0209] Fifteen thousand cells are plated into each well of a collagenI-coated 96-well plate in the DMEM growth medium. Six hours later, thecells are washed and the medium is replaced with DMEM without serum.After overnight culture to quiesce the cells, the medium is replaced byDulbecco's phosphate-buffered saline (Life Technologies Inc., GrandIsland, N.Y.) with 0.1% bovine albumin (Sigma Chemical Co., St Louis,Mo.). After adding various concentrations (0-300 nM) of test compoundsto the cells in 1% final concentration of DMSO, the cells are incubatedat room temperature for 30 minutes. The cells are then treated with VEGF(30 ng/ml) for 10 minutes at room temperature. Following VEGFstimulation, the buffer is removed and the cells are lysed by additionof 150 μl of extraction buffer (50 mM Tris, pH 7.8, supplemented with10% glycerol, 50 mM BGP, 2 mM EDTA, 10 mM NaF, 0.5 mM NaVO4, and 0.3%TX-100) at 4° C. for 30 minutes.

[0210] To assess receptor phosphorylation, 100 microliters of each celllysate is added to the wells of an ELISA plate precoated with 300 ng ofantibody C20 (Santa Cruz Biotechnology, Inc., Santa Cruz, Calif.).Following a 60-minute incubation, the plate is washed and bound KDR isprobed for phosphotyrosine using an anti-phosphotyrosine mAb clone 4G10(Upstate Biotechnology, Lake Placid, N.Y.). The plate is washed andwells are incubated with anti-mouse IgG/HRP conjugate (AmershamInternational plc, Buckinghamshire, England) for 60 minutes. Wells arewashed and phosphotyrosine is quantitated by addition of 100 μl per wellof 3,3′,5,5′ tetramethylbenzidine (Kirkegaard and Perry, TMB Microwell 1Component peroxidase substrate) solution. Color development is arrestedby the addition of 100 μl 1% HCl based stop solution (Kirkegaard andPerry, TMB 1 Component Stop Solution).

[0211] Optical densities (OD) are determined spectrophotometrically at450 nm in a 96-well plate reader (SpectraMax 250, Molecular Devices).Background (no VEGF added) OD values are subtracted from all ODs andpercent inhibition is calculated according to the equation:${\% \quad {Inhibition}} = \frac{( {{{OD}( {{VEGF}\quad {control}} )} - {{OD}( {{with}\quad {test}\quad {compound}} )}} ) \times 100}{{{OD}( {{VEGF}\quad {control}} )} - {{OD}( {{no}\quad {VEGF}\quad {added}} )}}$

[0212] IC₅₀s are determined on some of the exemplary materials with aleast squares analysis program using compound concentration versuspercent inhibition.

[0213] In vivo Assay of VEGFR Inhibition: Matrigel® Angiogenesis Model

[0214] Preparation of Matrigel Plugs and in vivo Phase: Matrigel®(Collaborative Biomedical Products, Bedford, Mass.) is a basementmembrane extract from a murine tumor composed primarily of laminin,collagen IV and heparan sulfate proteoglycan. It is provided as asterile liquid at 4° C., but rapidly forms a solid gel at 37° C.

[0215] Liquid Matrigel at 4° C is mixed with SK-MEL2 human tumor cellsthat are transfected with a plasmid containing the murine VEGF gene witha selectable marker. Tumor cells are grown in vitro under selection andcells are mixed with cold liquid Matrigel at a ratio of 2×10⁶ per 0.5ml. One half milliliter is implanted subcutaneously near the abdominalmidline using a 25 gauge needle. Test compounds are dosed as solutionsin Ethanol/Cremaphor EL/saline (12.5%:12.5%:75%) at 30, 100, and 300mg/kg po once daily starting on the day of implantation. Mice areeuthanized 12 days post-implantation and the Matrigel pellets areharvested for analysis of hemoglobin content.

[0216] Hemoglobin Assay: the Matrigel pellets are placed in 4 volumes(w/v) of 4° C. Lysis Buffer (20 mM Tris pH 7.5, 1 mM EGTA, 1 mM EDTA, 1%Triton X-100 [EM Science, Gibbstown, N.J.], and complete, EDTA-freeprotease inhibitor cocktail [Mannheim, Germany]), and homogenized at 4°C. Homogenates are incubated on ice for 30 minutes with shaking andcentrifuged at 14K×g for 30 minutes at 4° C. Supernatants aretransferred to chilled microfuge tubes and stored at 4° C for hemoglobinassay. Mouse hemoglobin (Sigma Chemical Co., St. Louis, Mo.) issuspended in autoclaved water (BioWhittaker, Inc, Walkersville, Md.) at5 mg/ml. A standard curve is generated from 500 micrograms/ml to 30micrograms/ml in Lysis Buffer (see above). Standard curve and lysatesamples are added at 5 microliters /well in duplicate to a polystyrene96-well plate. Using the Sigma Plasma Hemoglobin Kit (Sigma ChemicalCo., St. Louis, Mo.), TMB substrate is reconstituted in 50 mls roomtemperature acetic acid solution. One hundred microliters of substrateis added to each well, followed by 100 microliters/well of HydrogenPeroxide Solution at room temperature. The plate is incubated at roomtemperature for 10 minutes.

[0217] Optical densities are determined spectrophotometrically at 600 nmin a 96-well plate reader, SpectraMax 250 Microplate SpectrophotometerSystem (Molecular Devices, Sunnyvale, Calif.). Background Lysis Bufferreadings are subtracted from all wells. Total sample hemoglobin contentis calculated according to the following equation:

Total Hemoglobin=(Sample Lysate Volume)×(Hemoglobin Concentration)

[0218] The average Total Hemoglobin of Matrigel samples without cells issubtracted from each Total Hemoglobin Matrigel sample with cells.Percent inhibition is calculated according to the following equation:${\% \quad {Inhibition}} = \frac{( {{Average}\quad {Total}\quad {Hemoglobin}\quad {Drug}\text{-}{Treated}\quad {Tumor}\quad {Lysates}} ) \times 100}{( {{Average}\quad {Total}\quad {Hemoglobin}\quad {Non}\text{-}{Treated}\quad {Tumor}\quad {Lysates}} )}$

[0219] The three preceding assays establish that the compounds ofFormula I are active to inhibit VEGF receptor kinase activity and toinhibit angiogenesis.

[0220] In Vivo Assay of Antitumor Activity

[0221] An in vivo assay of the inhibitory effect of the compounds ontumors (e.g., solid cancers) mediated by raf kinase can be performed asfollows: CDI nu/nu mice (6-8 weeks old) are injected subcutaneously intothe flank at 1×10⁶ cells with human colon adenocarcinoma cell line. Themice are dosed i.p., i.v. or p.o. at 10, 30, 100, or 300 mg/Kg beginningon approximately day 10, when tumor size is between 50-100 mg. Animalsare dosed for 14 consecutive days; tumor size is monitored with caliperstwice a week. The inhibitory effect of the compounds on p38, raf andVEGFR kinases and therefore on tumor growth (e.g., solid cancers) canfurther be demonstrated in vivo according to the technique of Monia etal. (Nat. Med. 1996, 2, 668-75).

[0222] The preceding examples can be repeated with similar success bysubstituting the generically or specifically described reactants and/oroperating conditions of this invention for those used in the precedingexamples.

[0223] From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various conditions andusages.

What is claimed is:
 1. A compound of formula (I),

Wherein, Y is OR¹ or NHR², Hal is chlorine or bromine, R¹ is H or C₁-C₆alkyl R² is H, OH, CH₃ or CH₂OH, Z¹ and Z² are each H or OH, whereinonly one of Z¹ or Z² can be OH. X¹ to X⁷ are each, independently, H, OHor O(CO)C₁-C₄ alkyl, and n is 0 or 1, with the proviso that at least oneof conditions a-c is met, a) Z¹ or Z² is OH, b) Y is NHR² and R² is OH,c) n is 1, or a salt thereof, or an isolated stereoisomer thereof.
 2. Acompound of claim 1 wherein n of formula I is
 1. 3. A compound of claim2 wherein Y is NHR² and R² is H or CH₃,
 4. A compound of claim 2 whereina) X¹ to X⁷ are each H, or b) Z¹ and Z² are each H.
 5. A compound ofclaim 2 wherein a) X¹ to X⁷ are each H, or b) Z¹ is H and Z² is OH or Z¹is OH and Z² is H, or c) X¹ to X⁷ and Z¹ are each H and Z² is OH or d)X¹ to X⁷ and Z² are each H and Z¹ is OH.
 6. A compound of claim 2,wherein at least one of X¹ to X⁷ is OH or O(CO)C₁-C₄ alkyl.
 7. Acompound of claim 2, wherein Y is NHR² and R² is CH₂OH or OH.
 8. Acompound of claim 2 wherein Y is OH.
 9. A compound of claim 1, whereinZ¹ is H and Z² is OH or Z¹ is OH and Z² is H.
 10. A compound of claim 9,wherein n is
 0. 11. A compound of claim 10, wherein R² is H or CH₃. 12.A compound of claim 10, wherein X¹ to X⁷ are each H.
 13. A compound ofclaim 10, wherein at least one of X¹ to X⁷ is OH or O(CO)C₁-C₄ alkyl.14. A compound of claim 10, wherein R² is CH₂OH or OH.
 15. A compound ofclaim 10, wherein Y is OH.
 16. A compound of claim 1, wherein in formula(I), Y is NHR² and R² is OH.
 17. A compound of claim 16, wherein n is 0.18. A compound of claim 17, wherein X¹ to X⁷ are each H,
 19. A compoundof claim 17, wherein Z¹ is H and Z² is OH or Z¹ is OH and Z²is H.
 20. Acompound of claim 17, wherein at least one of X¹ to X⁷ is OH orO(CO)C₁-C₄ alkyl.
 21. A compound of claim 1, wherein in formula (I), Yis OH.
 22. A compound of claim 21, wherein n is
 0. 23. A compound ofclaim 22, wherein X¹ to X⁷ are each H.
 24. A compound of claim 22,wherein Z² is H and Z¹ is OH.
 25. A compound of claim 22, wherein Z¹ isH and Z² is OH.
 26. A compound of claim 22, wherein at least one of X¹to X⁷ is OH or O(CO)C₁-C₄ alkyl.
 27. A compound selected from the groupconsisting of:4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-hydroxymethyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-hydroxymethyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridinecarboxamide 1-oxide, salts thereof and stereoisomers thereof.
 28. Acompound of formula (II), or a salt or stereoisomer thereof,

wherein, Y is OR¹ or NHR², Hal is chlorine or bromine, R¹ is H or C₁-C₆alkyl R² is H, OH, CH₃ or CH₂OH, Z¹ and Z² are each H or OH, whereinonly one of Z¹ or Z² is OH, X⁴ to X⁷ are each, independently, H, OH orO(CO)C₁-C₄ alkyl, and n is 0 or 1, with the proviso that at least one ofconditions a-c is met, a) Z¹ or Z² is OH, b) Y is NHR² and R² is OH, c)n is
 1. 29. A compound of claim 28, wherein in formula (II), n is
 1. 30.A compound of claim 29, wherein in formula (II), Z¹ and Z² are each H.31. A compound of claim 30, wherein in formula (II), at least one of X⁴to X⁷ is OH.
 32. A compound of claim 30, wherein in formula (II), Y isNHR² and R² is H or CH₃.
 33. A compound of claim 28, wherein in formula(II), n is 0 and Z¹ is H and Z² is OH or Z¹ is OH and Z² is H.
 34. Acompound of claim 28, wherein in formula (II), n is 0, Z¹ and Z² areeach H, and at least one of X⁴ to X⁷ is OH.
 35. A compound of claim 33,wherein in formula (II), at least one of X⁴ to X⁷ is OH.
 36. A compoundof claim 33, wherein in formula (II), Y is NHR² and R² is H or CH₃. 37.A compound of claim 33, wherein in formula (II) Y is NHR² and R² is OH.38 A compound of claim 37, wherein in formula (II), at least one of X⁴to X⁷ is OH.
 39. A compound of formula (III), or a salt or isolatedstereoisomer thereof,

wherein, Y is OR¹ or NHR², Hal is chlorine or bromine, R¹ is H or C₁-C₆alkyl R² is H, OH, CH₃ or CH₂OH, Z¹ and Z² are each H or OH, whereinonly one of Z¹ or Z² can be OH, and n is 0 or 1, with the proviso thatat least one of conditions a-c is met, a) Z¹ or Z² is OH, b) Y is NHR²and R² is OH, c) n is
 1. 40. A compound of claim 39, wherein in formula(III), n is 1 and Z¹ and Z² are each H,
 41. A compound of claim 40,wherein in formula (III), Y is NHR² and R² is H or CH₃,
 42. A compoundof claim 39, wherein in formula (III), n is 0 and Z¹ is H and Z² is OHor Z¹ is OH and Z² is H,
 43. A compound of claim 42, wherein in formula(III), Y is NHR² and R² is H or CH₃.
 44. A compound of claim 39, whereinin formula (III), Y is OH.
 45. A method of preparing compounds of claim1 comprising the oxidation of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide, or 4-{4-[({[4-chloro-3-trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide.
 46. Amethod as in claim 45 wherein oxidation of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide, 4-{4-[({[4-chloro-3-trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide: a) replacesone or more of the phenyl hydrogens at the positions represented by X¹to X⁷ with a hydroxyl group, b) hydroxylates the N-methyl amide into ahydroxymethyl amide or hydroxamic acid, c) demethylates the N-methylamide into an unsubstituted amide, d) replaces one or more of the ureanitrogens (═NH) with a hydroxyl group to form an N-hydroxyurea (═NOH),e) hydrolyzes the N-methyl amide into a carboxylic acid, f) oxidizes thepyridyl ring nitrogen to form the corresponding pyridine-1-oxide, or g)provides a combination of two or more of a)-f); with the proviso that atleast one of b), d) and f) is performed.
 47. A method as in claim 46wherein oxidation of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide, 4-{4-[({[4-chloro-3-trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide, or4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-2-pyridine carboxamide replaces oneor more hydrogens at the positions represented by X¹ to X⁷ with ahydroxyl group and at least one of the hydroxyl groups in the X¹ to X⁷positions is esterified.
 48. A method as in claim 46 which prepares4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide, or a pharmaceutically acceptable salt of one ofthese oxides, or an isolated stereoisomer of one of these oxides.
 49. Apharmaceutical composition comprising an effective amount of at leastone compound of claim 1 and a physiologically acceptable carrier.
 50. Apharmaceutical composition comprising an effective amount of4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}-N-methyl-2-pyridinecarboxamide 1-oxide,4-{4-[({[4-chloro-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide,4-{4-[({[4-bromo-3-(trifluoromethyl)phenyl]amino}carbonyl)amino]phenoxy}2-pyridinecarboxamide 1-oxide or a pharmaceutically acceptable salt of one ofthese oxides, an isolated stereoisomer of one of these oxides or amixture thereof and a physiologically acceptable carrier.
 51. A methodof treating or preventing osteoporosis, inflammation, and angiogenesisdisorders, with the exclusion of cancer, in a mammal by administering aneffective amount of a compound of claim 1 to said mammal.
 52. A methodas in claim 51 wherein the compound of claim 1 administered is within apharmaceutical composition comprising an effective amount of a compoundof claim 1 and a physiologically acceptable carrier.
 53. A method oftreating or preventing a hyper-proliferative disorder in a mammalcomprising administering an effective amount of a compound of claim 1 tosaid mammal.
 54. A method of treating or preventing ahyper-proliferative disorder in a mammal comprising administering aneffective amount of a compound of claim 27 to said mammal.
 55. A methodof treating or preventing a hyper-proliferative disorder in a mammalcomprising administering to said mammal a) an effective amount of acompound of claim 1 and b) an additional anti-proliferative agent.
 56. Amethod as in claim 55 wherein the compound of claim 1 administered iswithin a pharmaceutical composition comprising an effective amount of acompound of claim 1 and a physiologically acceptable carrier.
 57. Amethod as in claim 56 wherein the pharmaceutical composition comprisesan effective amount of a compound of claim 1, a physiologicallyacceptable carrier and the additional anti-proliferative agent.
 58. Amethod as in claim 56 wherein the additional anti-proliferative agentadministered is within a pharmaceutical composition separate from thepharmaceutical composition comprising an effective amount of a compoundof claim 1 and a physiologically acceptable carrier.
 59. A method as inclaim 56 wherein the additional anti-proliferative agent is selectedfrom the group consisting of asparaginase, bleomycin, carboplatin,carmustine, chlorambucil, cisplatin, colaspase, cyclophosphamide,cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin(adriamycine), epirubicin, etoposide, 5-fluorouracil,hexamethylmelamine, hydroxyurea, ifosfamide, irinotecan, leucovorin,lomustine, mechlorethamine, 6-mercaptopurine, mesna, methotrexate,mitomycin C, mitoxantrone, prednisolone, prednisone, procarbazine,raloxifen, streptozocin, tamoxifen, thioguanine, topotecan, vinblastine,vincristine, vindesine, aminoglutethimide, L-asparaginase, azathioprine,5-azacytidine cladribine, busulfan, diethylstilbestrol, 2′,2′-difluorodeoxycytidine, docetaxel, erythrohydroxynonyladenine, ethinylestradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridine monophosphate,fludarabine phosphate, fluoxymesterone, flutamide, hydroxyprogesteronecaproate, idarubicin, interferon, medroxyprogesterone acetate, megestrolacetate, melphalan, mitotane, paclitaxel, oxaliplatin, gemcitabone,gefinitib, taxotere, BCNU, CCNU, DTIC, ara A, ara C, herceptin,actinomycin D, pentostatin, N-phosphonoacetyl-L-aspartate (PALA),plicamycin, semustine, teniposide, testosterone propionate, thiotepa,trimethylmelamine, uridine, and vinorelbine.
 60. A method of treating orpreventing osteoporosis, inflammation, and angiogenesis disorders, withthe exclusion of raf-mediated cancer, in a mammal by administering aneffective amount of a compound of claim 27 to said mammal.
 61. A methodof treating or preventing cancer by administering to a mammal a) aneffective amount of a compound of claim 1, and b) a cytotoxic agent orcytostatic chemotherapeutic agent.
 62. A method of claim 61 wherein thecompound of claim 1 administered is within a pharmaceutical compositioncomprising an effective amount of a compound of claim 1 and aphysiologically acceptable carrier.
 63. A method of claim 62 wherein thepharmaceutical composition comprises an effective amount of a compoundof claim 1, a physiologically acceptable carrier and the cytotoxic agentor cytostatic chemotherapeutic agent.
 64. A method of claim 62 whereinthe cytotoxic agent or cytostatic chemotherapeutic agent administered iswithin a pharmaceutical composition separate from the pharmaceuticalcomposition comprising an effective amount of a compound of claim 1 anda physiologically acceptable carrier.
 65. A method as in claim 61wherein the cytotoxic or cytostatic chemotherapeutic agent is selectedfrom the group consisting of DNA topoisomerase I and II inhibitors, DNAintercalators, alkylating agents, microtubule disruptors, hormone andgrowth factor receptor agonists or antagonists, other kinase inhibitorsand anti-metabolites.
 66. A kit comprising a separate dose of thecytotoxic or cytostatic agent and, a separate dose of a compound ofclaim
 1. 67. A method of treating or preventing a hyper-proliferativedisorder in a mammal comprising administering to said mammal a) aneffective amount of a compound of claim 27 and b) an additionalanti-proliferative agent.
 68. A method as in claim 67 wherein thecompound of claim 27 administered is within a pharmaceutical compositioncomprising an effective amount of a compound of claim 27 and aphysiologically acceptable carrier.
 69. A method as in claim 68 whereinthe pharmaceutical composition comprises an effective amount of acompound of claim 27, a physiologically acceptable carrier and theadditional anti-proliferative agent.
 70. A method as in claim 68 whereinthe additional anti-proliferative agent administered is within apharmaceutical composition separate from the pharmaceutical compositioncomprising an effective amount of a compound of claim 27 and aphysiologically acceptable carrier.
 71. A method as in claim 68 whereinthe additional anti-proliferative agent is selected from the groupconsisting of asparaginase, bleomycin, carboplatin, carmustine,chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine,dacarbazine, dactinomycin, daunorubicin, doxorubicin (adriamycine),epirubicin, etoposide, 5-fluorouracil, hexamethylmelamine, hydroxyurea,ifosfamide, irinotecan, leucovorin, lomustine, mechlorethamine,6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone,prednisolone, prednisone, procarbazine, raloxifen, streptozocin,tamoxifen, thioguanine, topotecan, vinblastine, vincristine, vindesine,aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidinecladribine, busulfan, diethylstilbestrol, 2′,2′-difluorodeoxycytidine,docetaxel, erythrohydroxynonyladenine, ethinyl estradiol,5-fluorodeoxyuridine, 5-fluorodeoxyuridine monophosphate, fludarabinephosphate, fluoxymesterone, flutamide, hydroxyprogesterone caproate,idarubicin, interferon, medroxyprogesterone acetate, megestrol acetate,melphalan, mitotane, paclitaxel, oxaliplatin, gemcitabone, gefinitib,taxotere, BCNU, CCNU, DTIC, ara A, ara C, herceptin, actinomycin D,pentostatin, N-phosphonoacetyl-L-aspartate (PALA), plicamycin,semustine, teniposide, testosterone propionate, thiotepa,trimethylmelamine, uridine, and vinorelbine.
 72. A method of preparingN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)-1-oxo-(4-pyridyloxy)]phenyl}urea

comprising chemically oxidizingN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-(N-methylcarbamoyl)(4-pyridyloxy)]phenyl}ureain solution.
 73. A method of preparingN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-1-oxo-(4-pyridyloxy)]phenyl}urea

comprising chemically oxidizingN-[4-chloro-3-(trifluoromethyl)phenyl]-N′-{4-[2-carbamoyl-(4-pyridyloxy)]phenyl}ureain solution.